Alkoxythiophene formyl arylamine compound and application thereof

Abstract

The invention provides an alkoxy thiophene formyl arylamine compound and application thereof. The alkoxy thiophene formyl arylamine compound comprises the alkoxy thiophene formyl arylamine compound with the structure shown in the formula (1) or pharmaceutically acceptable salt of the alkoxy thiophene formyl arylamine compound. The alkoxy thiophene formyl arylamine compound provided by the invention is a sphingomyelin synthase inhibitor with a novel structure, has remarkable inhibitory activity on SMS2, ideal physicochemical properties such as stability and water solubility, and small potential toxic and side effects, and can be further prepared into drugs for preventing or treating diseases related to abnormal sphingomyelin level. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of medicinal chemistry, and more specifically, relates to sphingomyelin synthase inhibitor alkoxythiophene carboxylic acid aromatic amide compounds and applications thereof. Background Art

[0002] Atherosclerosis (AS) is one of the main pathological bases of many cardiovascular and cerebrovascular diseases. Therefore, the research on anti-atherosclerotic drugs has become a hot topic in the current field of drug research and development.

[0003] Studies have proposed a variety of potential drug targets for anti-atherosclerosis, such as sphingomyelin synthase inhibitors, PPAR agonists, apolipoprotein infusion, liver X receptor activators, and phospholipid transfer protein (PLTP) inhibitors. In particular, sphingomyelin (SM) and its metabolic enzymes mediate a series of cellular processes while causing lipoprotein changes, indicating that they play an important role in the development of atherosclerosis. The industry believes that reducing the level of sphingomyelin in plasma or inhibiting the synthesis of SM can achieve the purpose of slowing down or preventing the development of atherosclerosis.

[0004] In addition, researchers have found that sphingomyelin synthase (SMS) can regulate the synthesis of SM from ceramide and phosphatidylcholine (PC), and is a key enzyme in the last step of the de novo synthesis pathway of sphingomyelin. There are three main subtypes of sphingomyelin synthase, sphingomyelin synthase 1 (SMS1), sphingomyelin synthase 2 (SMS2) and sphingomyelin synthase-related protein (SMSr). Studies have shown that reducing sphingomyelin levels by inhibiting SMS2 can become a new method for treating atherosclerosis. SMS2 has potential advantages as a new target for anti-atherosclerosis, and SMS2 selective inhibitors may become new anti-atherosclerotic therapeutic drugs. In addition, studies have found that SMS2 deficiency can prevent high-fat diet-induced obesity and insulin resistance. At the same time, it is difficult to observe large mature fat plaques in the liver of SMS2 knockout mice, indicating that SMS2 is involved in the formation of liver fat plaques and can induce obesity and type II diabetes (Mitsutake et al., 2011). Small molecule inhibitors of sphingomyelin synthase may be used to prevent and treat metabolic syndromes such as type II diabetes, obesity and fatty liver. Literature reports that SMS2 gene knockout can significantly improve inflammation and insulin resistance and other metabolic syndromes in mice fed with high fat (Mitsutake et al., 2011). SMS2 small molecule inhibitors may be used to prevent and treat inflammatory-related diseases such as colitis and colitis-related diseases, colon cancer, pulmonary edema and lung injury. Studies have found that SMS2 can promote the EMT process of breast cancer cells (Zheng et al., 2019), drug resistance of colon cancer cells (Jin et al., 2016), growth and metastasis of ovarian cancer (Jing et al., 2021), M2 polarization of macrophages in the tumor microenvironment of breast cancer (Deng et al., 2020), infiltration of malignant lymphoma (Taniguchi et al., 2020) and growth of leukemia cells (JP2021-165292A; WO2005032492), etc. Small molecule inhibitors of sphingomyelin synthase may be used to prevent and treat malignant tumors, such as triple-negative breast cancer, colon cancer, leukemia, ovarian cancer, etc. The loss of SMS2 can inhibit the content and aggregation of sphingomyelin in the brain and reduce the content of Aβ (Kidani et al., 2012; Yuyama et al., 2014; Hsiao et al., 2013). Small molecule inhibitors of SMS2 may be used to prevent and treat central nervous system diseases, such as neurodegenerative diseases such as Alzheimer's disease.Literature has revealed that there are some lipid abnormalities in the tears of dry eye, including sphingomyelin (Ham et al., 2006; Paranjpe et al., 2019), suggesting that SMS2 inhibitors can be used to prevent and alleviate diseases related to abnormal sphingomyelin metabolism such as dry eye.

[0005] With the deepening of the study of SMS biological function, people have found that SMS2 is related to the occurrence and development of many diseases. In recent years, a variety of SMS inhibitors have been discovered and some have been applied to the evaluation of certain disease treatment models. Aimin Meng et al. used a selective tumor cytotoxic agent, D609, to treat U937 human monocytic leukemia cells, and detected the levels of intracellular ceramide and DAG to measure the degree of SMS inhibition. They found that D609-induced cell death was time- and concentration-dependent, accompanied by high levels of intracellular ceramide and low levels of SM and DAG; using PKC inhibitors and agonists to treat cells with D609, they found that PKC inhibitors could increase cell apoptosis, while agonists had the opposite effect, thus proving to a certain extent that SMS is a potential target of D609 (Meng et al., 2004). Salma Y et al. found that Jaspine B isolated from sponges could significantly inhibit the SMS activity of melanoma cells and induce apoptosis of melanoma cells (Salma et al., 2009). Mahadeva MMSwamy et al. designed and synthesized a series of ceramide analogs, such as compound 17j, which have a certain SMS inhibitory effect (Swamy et al., 2018). Muhamad Aqmal Othman et al. found that Malabaricone C, a compound isolated from the fruit of Myristica cinnamomea King, can inhibit SMS2, but has low selectivity for SMS2 and SMS1, and can significantly reduce the weight gain, glucose tolerance and liver fatty degeneration levels of HFD-induced mice (Swamy et al., 2018). Ryutaro Adachi et al. established a radioactive hydrogen substrate labeled SMS activity measurement system, and obtained a class of quinolone compounds (such as compound 3) with good SMS2 inhibitory activity and selectivity through high-throughput screening and structural optimization (Adachi et al., 2017); similarly, Takafumi Yukawa et al. obtained a class of 1,8-naphthylidene-2-one compounds, such as compound 37, by modifying the above-mentioned quinolone compounds (Yukawa et al., 2020). Xiaodong Deng et al. screened and designed a new class of 2-benzyloxyphenylacetonitrile derivatives through virtual screening and enzyme activity assay, which have preliminary SMS2 inhibitory activity, such as D2 (Deng et al., 2014). Ya-li Li et al. obtained a class of SAPA series compounds with good inhibitory activity against SMS1 through virtual screening, such as SAPA1a (Li et al., 2015).Xiang-Yu Qi et al. synthesized a class of oxazolopyridine derivatives with good SMS2 inhibitory activity and selectivity through skeleton hopping combined with molecular docking design (Qi et al., 2017), such as QY16. Mingguang Mo et al. obtained a class of benzisoxazole derivatives through skeleton hopping and ring closure, and found that 15w in this class of compounds has good SMS2 inhibitory activity and selectivity and good pharmacokinetic properties. At an oral dose of 50 mg / kg, it can significantly inhibit the level of inflammatory factors in db / db mice, reduce LDL-C and increase HDL-C levels (Mo et al., 2018). Yali Li et al. designed and obtained a class of salicylamide derivatives by means of electron isosteres, and found that Ly93 in this class of compounds has high SMS2 inhibitory activity and selectivity, which can inhibit liver SMS activity in a dose-dependent manner, reduce plasma SM levels, and ultimately inhibit the development of atherosclerosis; in vitro test results showed that Ly93 reduced apoB secretion of Huh7 cells in a dose-dependent manner, reduced SMS activity of macrophages, and increased cholesterol efflux; at the same time, Ly93 can inhibit LPS-mediated secretion of macrophage proinflammatory factors and chemokines (Li et al., 2019). Hadya Virupaksha Deepak et al. found that Daurichromenic acid (DCA), a compound isolated from Rhododendrondauricum plants, can inhibit SMS activity and show inhibition of Aβ aggregation (EC. 50 =57 μM) (Deepak et al., 2020). In general, although many SMS inhibitors have been discovered, the structural type and drugability of SMS2 inhibitors need to be further optimized. Summary of the invention

[0006] The first object of the present invention is to provide an alkoxythiophene carboxylic acid aromatic amide compound (including carboxyl / phosphate compounds derived therefrom as prodrugs) which has significant inhibitory activity against SMS2 and ideal physicochemical properties such as stability and water solubility.

[0007] The second object of the present invention is to provide a pharmaceutical composition containing alkoxythiophene carboxylic acid aromatic amide compounds (including carboxylic acid / phosphate compounds derived therefrom as prodrugs) or pharmaceutically acceptable salts thereof.

[0008] The third object of the present invention is to provide pharmaceutical uses of alkoxythiophene carboxylic acid aromatic amide compounds (including carboxylic acid / phosphate compounds derived therefrom as prodrugs) or pharmaceutically acceptable salts thereof.

[0009] In order to achieve the above object, the present invention provides an alkoxythiophene carboxylic acid aromatic amide compound having a structure shown in formula (1), or a pharmaceutically acceptable salt thereof:

[0010]

[0011] Wherein, X, Y, and Z are each independently selected from a carbon atom or a sulfur atom;

[0012] R 1 is selected from substituted or unsubstituted phenyl, pyridyl, pyridazinyl, pyrimidinyl, thiazolyl, wherein the substituents of the substituted groups are selected from 1 to 5 substituents of halogen, methyl, hydroxy, methoxy, trifluoromethyl, trifluoromethoxy, cyano, and nitro;

[0013] R 2 Selected from hydrogen, alkyl with less than 3 carbon atoms, and cycloalkyl;

[0014] R 3 is selected from substituted or unsubstituted hydrocarbon groups;

[0015] R 4 One or two selected from hydrogen, alkyl with less than 3 carbon atoms, halogen, and nitro.

[0016] As a preferred embodiment, X and Y are each independently selected from a sulfur atom or a carbon atom, and Z is a carbon atom.

[0017] As a preferred solution, the R 3 Select one of the following structures:

[0018]

[0019] Wherein n is an integer from 1 to 10;

[0020] W is selected from a carbon atom or a nitrogen atom;

[0021] L, M are each independently selected from -C=C-, a sulfur atom, an oxygen atom;

[0022] R 5 1 to 5 selected from hydrogen, substituted or unsubstituted alkyl with less than 4 carbon atoms, substituted or unsubstituted alkoxy with less than 20 carbon atoms, halogen, acyl, nitro, cyano, hydroxyl, carboxylic (phosphonic) ester, and aromatic heterocycle.

[0023] As a preferred embodiment, the alkoxy group is an alkoxy group with less than 15 carbon atoms.

[0024] As a preferred embodiment, the compound is:

[0025]

[0026]

[0027]

[0028]

[0029] As a preferred embodiment, the pharmaceutically acceptable salt is hydrochloride, hydrobromide, tartrate, methanesulfonate, sodium salt, potassium salt, calcium salt, ammonium salt or amino acid salt.

[0030] The compounds of the present invention contain basic groups that can form salts with acids, and salts of derivatives can be formed by common means, including organic acid salts such as acetates, citrates, fumarates, maleates, oxalates, malates, citrates, succinates, tartrates, lactates, camphorsulfonates, benzenesulfonates, p-toluenesulfonates, methanesulfonates, trifluoroacetates, trifluoromethanesulfonates, etc.; inorganic acid salts such as hydrohalic acid (hydrofluoric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid) salts, sulfates, phosphates, nitrates, etc. Or with amino acids, such as glutamic acid or aspartic acid, glutamate or aspartate can be formed, and the preferred salts are hydrochloride and hydrobromide; similarly, some compounds of the present invention contain acidic groups that can form salts with bases, and salts of derivatives can be formed by common means, including sodium salts, potassium salts, calcium salts, ammonium salts, or with amino acids such as lysine, arginine, histidine, etc., and the preferred salts are sodium salts and potassium salts.

[0031] In order to achieve the second object of the present invention, the present invention provides a pharmaceutical composition comprising at least one therapeutically effective amount of an alkoxythiophenecarboxylic acid aromatic amide compound represented by formula (1), or a pharmaceutically acceptable salt thereof.

[0032] The pharmaceutical composition of the present invention may also contain one or more pharmaceutically acceptable carriers, including conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field, and flavoring agents, sweeteners, etc. may also be added when necessary.

[0033] The pharmaceutical composition of the present invention can be prepared in any form, such as granules, powders, tablets, coated tablets, capsules, pills, syrups, drops, solutions, suspensions and emulsions, or sustained-release preparations of the active ingredients, wherein capsules include hard or soft gelatin capsules, and granules and powders can be non-effervescent or effervescent forms.

[0034] The pharmaceutical composition of the present invention can be administered by various routes according to conventional methods, including oral, intravenous, intraarterial, intraperitoneal, intrathoracic, transdermal, nasal, inhalation, rectal, ocular and subcutaneous administration.

[0035] In order to achieve the third object of the present invention, the present invention provides the use of the alkoxythiophene carboxylic acid aromatic amide compound or a pharmaceutically acceptable salt thereof in the preparation of a sphingomyelin synthase small molecule inhibitor.

[0036] The present invention adopts the high performance liquid chromatography (HPLC) fluorescence quantitative detection method reported in the literature to determine the inhibitory activity of the alkoxythiophene carboxylic acid aromatic amide compound shown in formula (1) on sphingomyelin synthase (Deng et al., 2012), and calculates the change in the activity of the inhibitor on sphingomyelin synthase catalyzing ceramide to sphingomyelin through the change in the content of NBD-ceramide and NBD-sphingomyelin. The activity test experiment based on the high performance liquid chromatography (HPLC) fluorescence quantitative method shows that the alkoxythiophene carboxylic acid aromatic amide compound shown in formula (1) (including the carboxyl / phosphate compound derived therefrom as a prodrug) has micro / submicromole SMS2 inhibitory activity, and the high performance liquid chromatography (HPLC) fluorescence quantitative method is used to detect the inhibitory activity of the compound on SMS2.

[0037] The present invention also provides the use of the alkoxythiophene carboxylic acid aromatic amide compound or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and treating diseases caused by abnormal increase in sphingomyelin levels.

[0038] As a preferred embodiment, the diseases caused by abnormal increase in sphingomyelin levels include atherosclerosis, obesity, insulin resistance, fatty liver, type II diabetes, enteritis, pulmonary edema, lung injury, malignant tumors, Alzheimer's disease and dry eye. The present invention also provides evaluation data on the therapeutic effect of the pharmaceutical composition of the alkoxythiophene carboxylic acid aromatic amide compound or a pharmaceutically acceptable salt thereof on dry eye in mice.

[0039] The present invention has confirmed through experiments that the disclosed compounds have significant inhibitory activity on SMS2; ideal physical and chemical properties such as stability and water solubility; and they do not contain potential toxic groups, and have small potential toxic and side effects. They can be used as drugs for preventing or treating diseases related to abnormal sphingomyelin levels, including metabolic syndrome related to metabolic disorders, such as atherosclerosis, obesity, insulin resistance, fatty liver, and type II diabetes; diseases related to inflammation, such as enteritis, pulmonary edema, and lung injury; malignant tumors, such as triple-negative breast cancer, colon cancer, leukemia, ovarian cancer, etc.; and central nervous system diseases, such as neurodegenerative diseases such as Alzheimer's disease, dry eye syndrome, etc.

[0040] The term "halogen" refers to fluorine, chlorine, bromine and iodine, with fluorine and chlorine being preferred.

[0041] The term "alkyl" includes saturated aliphatic groups, including straight chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc.), branched chain alkyl groups (isopropyl, tert-butyl, isobutyl, etc.), cycloalkyl groups (cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl), alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In certain embodiments, the backbone of a straight or branched alkyl group has 4 or fewer carbon atoms.

[0042] The term "alkoxy" includes substituted and unsubstituted alkyl groups covalently linked to an oxygen atom. In certain embodiments, a linear or branched alkoxy group has 20 or fewer carbon atoms in its backbone, more preferably 15 or fewer carbon atoms.

[0043] The beneficial effect of the present invention is that the alkoxythiophene carboxylic acid aromatic amide compounds provided by the present invention (including carboxyl / phosphate compounds derived therefrom as prodrugs) are a class of novel structural sphingomyelin synthase inhibitors, have significant inhibitory activity against SMS2, ideal physical and chemical properties such as stability and water solubility, do not contain potential toxic groups, have small potential toxic and side effects, and can be further prepared into drugs for preventing or treating diseases related to abnormal sphingomyelin levels.

[0044] Instruction Manual

[0045] Figure 1 . The fluorescein sodium staining scores of mouse eyes were significantly higher in the pathological groups (including vehicle, cyclosporine, 1-2 and 1-31 groups) compared with the normal mice (physiological control group), and both SMS2 inhibitors and cyclosporine in the former could reduce the pathological scores of dry eyes. *p<0.05, **p<0.01, ****p<0.0001.

[0046] Figure 2 .The mouse eyeball was stained with sodium fluorescein. The pathological groups (including vehicle control group, cyclosporine, 1-2 and 1-31 groups) had more green areas after staining compared with the normal mice (physiological control group), indicating that the degree of corneal damage also increased. Compared with the vehicle group, the SMS2 inhibitor cyclosporine could reduce the area of ​​corneal damage, and the effect of the former was more significant.

[0047] Figure 3 The tear secretion of mice was statistically analyzed. The cotton thread soaking length of the pathological groups (including vehicle, cyclosporine, 1-2 and 1-31 groups) was reduced compared with the normal mice (physiological control group), indicating that the tear secretion was reduced. Compared with the vehicle group, the SMS2 inhibitor cyclosporine and sporine could promote the tear secretion of mice, and the former was more significant, *p<0.05, ****p<0.0001. DETAILED DESCRIPTION

[0048] The technology of the present invention is described in detail below in conjunction with specific embodiments. It should be noted that the following specific embodiments are only used to help those skilled in the art understand the present invention, and are not intended to limit the present invention.

[0049] Example 1: Preparation of 3-benzyloxy-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-1)

[0050] 1) Synthesis of methyl 3-benzyloxythiophene-2-carboxylate

[0051]

[0052] Weigh 5g (0.03mol, 1.0eq) of compound 2, 13g (0.09mol, 3.0eq) of potassium carbonate, and 500mg of potassium iodide in a 250mL round-bottom flask, add 100mL of acetonitrile to dissolve, add 8.1g (0.045mol, 1.5eq) of compound 3 under stirring, stir overnight at room temperature, and the reaction is complete after TLC detection. Filter and evaporate to dryness to obtain 7.4g of yellow oil, with a yield of 94%, MS (ESI) (m / z): 249 [M+H] + It can be used in the next reaction without further purification.

[0053] 2) Synthesis of 3-Benzyloxythiophene-2-carboxylic acid

[0054]

[0055] Weigh 7 g (0.03 mol, 1.0 eq) of compound 4 into a 250 mL round-bottom flask and add a hydroxide solution (2 mol / L, H 2 O / dioxane=1 / 1)100mL, transferred to oil bath and refluxed for 8h, TLC detected that the reaction was complete, cooled, adjusted pH to neutral with dilute hydrochloric acid, extracted with EA 3 times, combined EA phase, washed once with saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and recrystallized with PE / EA=10 / 1 to obtain 6g of yellow crystals, with a yield of 85%, MS (ESI) (m / z): 233[MH] - .

[0056] 3) Synthesis of 3-benzyloxy-N-(pyridin-3-yl)thiophene-2-carboxamide (Compound 1-1) (General Synthesis Method 1)

[0057]

[0058] Weigh 3g (0.013mol, 1.0eq) of compound 5 and 7.3g (0.019mol, 1.5eq) of HBTU into a 100mL round-bottom flask, add 50mL of DMF to dissolve, add 4.95g (0.038mol, 3.0eq) of DIPEA under stirring, stir at room temperature for 10min, add 3.6g (0.038mol, 3.0eq) of compound 6, stir at room temperature overnight, TLC detection reaction is complete, EA dissolves, wash with saturated brine 3 times, dry with anhydrous sodium sulfate, and flash purify to obtain 3.6g of yellow solid, i.e. 3-benzyloxy-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-1), yield 89%. MS (ESI) (m / z): 311 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.47 (s, 1H), 8.58 (s, 1H), 8.28 (m, 1H), 8.04 (d, J = 8.2Hz, 1H), 7.8 6(d,J=5.5Hz,1H),7.59(d,J=7.1Hz,2H),7.41(m,4H),7.28(d,J=5.4Hz,1H),5.44(s,2H).

[0059] Example 2: Preparation of Compounds 1-4, 1-5, 1-6, 1-7, 1-9, 1-14, 1-15, 1-20, 1-40, 1-61, 1-64, 1-65, 1-68, 1-69, 1-83

[0060] Referring to the conditions and steps of the first synthetic method in Example 1, 3-aminopyridazine was reacted with compound 5 to obtain 3-benzyloxy-N-(pyridazin-3-yl)thiophene-2-carboxamide (Formula 1-4) with a yield of 50%. MS (ESI) (m / z): 312 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.29(d,1H),8.61(d,1H),7.76(d,1H),7.60(t,1H),7.47–7.35(m,4H),7.27(s,1H),6.59(m,1H),5.20(s,2H).

[0061] Referring to the conditions and steps of the first synthetic method in Example 1, 2-amino-4-trifluoromethylpyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(4-trifluoromethylpyridin-2-yl)thiophene-2-carboxamide (Formula 1-5) with a yield of 16%. MS (ESI) (m / z): 379 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.97 (s, 1H), 8.64 (s, 1H), 8.43 (s, 1H), 7.53–7.33 (m, 5H), 7.27 (s, 1H), 7.23 (s, 1H), 6.91 (d, J = 4.5Hz, 1H), 5.40 (s, 2H).

[0062] Referring to the conditions and steps of the first synthetic method in Example 1, 2-amino-4-chloropyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(4-chloropyridin-2-yl)thiophene-2-carboxamide (Formula 1-6) with a yield of 46%. MS (ESI) (m / z): 345, 347 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.84(s,1H),8.43(s,1H),8.18(s,1H),7.56–7.33(m,5H),7.27(s,1H),7.03(s,1H),6.89(s,1H),5.39(s,2H).

[0063] Referring to the conditions and steps of the first synthetic method in Example 1, aniline was reacted with compound 5 to obtain 3-benzyloxy-N-phenylthiophene-2-carboxamide (Formula 1-7) with a yield of 68%. MS (ESI) (m / z): 310 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.20(s,1H),7.49(m,5H),7.43(m,2H),7.28(s,2H),7.27–7.25(m,1H),7.07(s,1H),6.99(s,1H),5.29(s,2H).

[0064] Referring to the conditions and steps of the first synthetic method in Example 1, 2-amino-6-hydroxypyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(6-hydroxypyridin-2-yl)thiophene-2-carboxamide (Formula 1-9) with a yield of 48%. MS (ESI) (m / z): 327 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.53(s,1H),7.76(d,J=7.5Hz,1H),7.58(t,J=7.5Hz,1H),7.43(dd,J=7.5,1.5Hz,2H) ,7.37(m,2H),7.27(m,1H),7.03(dd,J=7.5,1.4Hz,1H),6.68–6.51(m,2H),6.51–6.45(m,1H),5.20(s,2H).

[0065] Referring to the conditions and steps of the first synthetic method in Example 1, 3-benzyloxy-5-nitrothiophene-2-carboxylic acid was reacted with compound 6 to obtain 3-benzyloxy-5-nitro-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-14) with a yield of 56%. MS (ESI) (m / z): 356 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.16(s,1H),8.34(s,1H),8.20(m,2H),7.78(s,1H),7.50(m,2H),7.25(m,4H),5.32(s,2H).

[0066] Referring to the conditions and steps of the first synthetic method in Example 1, 3-benzyloxy-4-bromothiophene-2-carboxylic acid was reacted with compound 6 to obtain 3-benzyloxy-4-bromo-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-15) with a yield of 32%. MS (ESI) (m / z): 388, 390 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.03(s,1H),8.29(s,1H),7.99(s,2H),7.55–7.39(m,5H),7.24(s,1H),7.19(s,1H),5.35(s,2H).

[0067] Referring to the conditions and steps of the first synthetic method in Example 1, 2-amino-4,6-dichloropyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(4,6-dichloropyridin-2-yl)thiophene-2-carboxamide (Formula 1-20) with a yield of 7%. MS (ESI) (m / z): 379,381 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.90 (s, 1H), 8.34 (s, 1H), 7.50 (d, J = 15.8Hz, 5H), 7.41 (s, 1H), 7.28 (s, 1H), 7.06 (s, 1H), 6.94 (s, 1H), 5.38 (s, 2H).

[0068] Referring to the conditions and steps of the first synthetic method in Example 1, 3-benzyloxy-5-methylthiophene-2-carboxylic acid was reacted with compound 6 to obtain 3-benzyloxy-5-methyl-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-40) with a yield of 55%. MS (ESI) (m / z): 325 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.14 (s, 1H), 8.22 (d, J = 29.9Hz, 3H), 7.48 (m, 5H), 7.20 (m, 1H), 6.71 (s, 1H), 5.23 (s, 2H), 2.51 (s, 3H).

[0069] Referring to the conditions and steps of the first synthetic method in Example 1, 2-aminopyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(pyridin-2-yl)thiophene-2-carboxamide (Formula 1-61) with a yield of 68%. MS (ESI) (m / z): 311 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.78 (s, 1H), 8.31 (d, J = 7.8Hz, 2H), 7.70 (s, 1H), 7.43 (m, 5H), 7.28 (s, 1H), 7.02 (s, 1H), 6.88 (s, 1H), 5.40 (s, 2H).

[0070] Referring to the conditions and steps of the first synthetic method in Example 1, 5-aminopyrimidine was reacted with compound 5 to obtain 3-benzyloxy-N-(pyrimidin-5-yl)thiophene-2-carboxamide (Formula 1-64) with a yield of 43%. MS (ESI) (m / z): 312 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.58(s,1H),8.98(m,2H),8.91(s,1H),7.89(s,1H),7.59(m,2H),7.42(m,3H),7.26(s,1H),5.45(s,2H).

[0071] Referring to the conditions and steps of the first synthetic method in Example 1, 5-aminothiazole was reacted with compound 5 to obtain 3-benzyloxy-N-(thiazol-5-yl)thiophene-2-carboxamide (Formula 1-65) with a yield of 48%. MS (ESI) (m / z): 317 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.42(s,1H),8.62(s,1H),7.86(s,1H),7.71(s,1H),7.55(m,2H),7.40(d,J=24.8Hz,3H),7.24(s,1H),5.45(s,2H).

[0072] Referring to the conditions and steps of the first synthetic method in Example 1, 2-methyl-3-aminopyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(2-methylpyridin-3-yl)thiophene-2-carboxamide (Formula 1-68) with a yield of 53%. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.05(s,1H),8.37(s,1H),8.15(s,1H),7.89(s,1H),7 .59(m,2H),7.43(m,3H),7.34(s,1H),7.21(s,1H),5.42(s,2H),1.96(s,3H).

[0073] Referring to the conditions and steps of the first synthetic method in Example 1, 3-amino-4-methylpyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(4-methylpyridin-3-yl)thiophene-2-carboxamide (Formula 1-69) with a yield of 53%. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.38 (s, 1H), 8.84 (s, 1H), 8.21 (s, 1H), 7.55–7.42 (m, 6H), 7.00 (d, J = 11.6Hz, 2H), 5.26 (s, 2H), 1.73 (s, 3H).

[0074] Referring to the conditions and steps of the first synthetic method in Example 1, 3-amino-5-methylpyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(5-methylpyridin-3-yl)thiophene-2-carboxamide (Formula 1-83) with a yield of 72%. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.41(s,1H),8.42(s,1H),8.13(s,1H),7.86(s,1H),7 .78(s,1H),7.60(m,2H),7.44(s,3H),7.28(s,1H),5.43(s,2H),2.28(s,3H).

[0075] Example 3: Preparation of 3-(2-chloro-5-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-2)

[0076] 1) Synthesis of 3-hydroxy-N-(pyridin-3-yl)thiophene-2-carboxamide

[0077]

[0078] 2.3 g of compound 1-1 was weighed into a 100 mL round-bottom flask, 30 mL of anhydrous dichloromethane was added to dissolve, 1 mL of boron tribromide was added under stirring, and the mixture was stirred overnight at room temperature to precipitate a large amount of yellow solid. The reaction was complete by TLC detection. An appropriate amount of methanol was added to quench the dissolution, evaporated to dryness, an appropriate amount of water was added, the pH was adjusted to alkaline with a saturated sodium carbonate solution, acidified with dilute hydrochloric acid, extracted with EA, dried over anhydrous sodium sulfate, and evaporated to obtain 1 g of a yellow solid with a yield of 65%. MS (ESI) (m / z): 221 [M+H] + This compound was used in the next reaction without further purification.

[0079] 2) Synthesis of 3-(2-chloro-5-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-2) (General Synthesis Method 2)

[0080]

[0081] 70 mg (0.32 mmol, 1.0 eq) of compound 7 and 88 mg (0.64 mmol, 2.0 eq) of potassium carbonate were weighed into a round-bottom flask, 3 mL of NMP was added to dissolve, 92 mg (0.413 mmol, 1.3 eq) of compound 8 was added under stirring, and the mixture was stirred overnight at room temperature. The reaction was complete after TLC detection, EA was added to dissolve, the mixture was washed with saturated brine 3 times, and dried over anhydrous sodium sulfate. After flash purification, 75 mg of a white solid was obtained, namely 3-(2-chloro-5-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-2), with a yield of 65%. MS (ESI) (m / z): 363, 365 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.62(s,1H),8.29(m,1H),8.05(s,1H),7.89(s,1H),7.65(m,2H),7.34(m,2H),5.48(s,2H).

[0082] Example 4: Compounds 1-3, 1-8, 1-10, 1-11, 1-12, 1-13, 1-16, 1-18, 1-19, 1-21, 1-22, 1-23, 1-25, 1-27, 1-28, 1-29, 1-30, 1-31, 1-32, 1-33, 1-34, 1-35, 1-36, 1-37, 1-38, 1-39, 1-41, 1-42, 1-43, 1-44, 1-45, 1-46, 1-47, 1-48, 1-49, 1-50, Preparation of 1-51, 1-52, 1-53, 1-54, 1-55, 1-56, 1-57, 1-58, 1-59, 1-60, 1-63, 1-66, 1-67, 1-70, 1-71, 1-72, 1-73, 1-74, 1-75, 1-76, 1-77, 1-78, 1-79, 1-80, 1-81, 1-82, 1-84, 1-85, 1-86, 1-87, 1-88, 1-89, 1-93, 1-94, 1-97, 1-98, 1-99

[0083] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2,6-dichlorobenzyl bromide was reacted with compound 7 to obtain 3-(2,6-dichlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-3) with a yield of 75%. MS (ESI) (m / z): 379,381 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.22(s,1H),8.36(d,J=2.3Hz,1H),8.27(d,J=4.6Hz,1H),7.95(d,J=5.6Hz,2H) ,7.64(s,1H),7.62(s,1H),7.53(m,1H),7.45(d,J=5.5Hz,1H),7.36(dd,J=8.2,4.7Hz,1H),5.60(s,2H).

[0084] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 5-fluoro-2-(2-hydroxy-2-methylpropoxy)benzyl bromide was reacted with compound 7 to obtain 3-(5-fluoro-2-(2-hydroxy-2-methylpropoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-8) with a yield of 44%. MS (ESI) (m / z): 416 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.98(s,1H),8.51(d,J=1.3Hz,1H),8.24(dd,J=7.5,1.3Hz,1H),8.06(d,J=7.5Hz,1H),7.77(d,J=7.3Hz,1H ),7.30–7.22(m,2H),7.10–7.01(m,2H),6.97(m,1H),6.59(d,J=7.5Hz,1H),5.21(s,2H),4.25(s,2H),2.19(s,1H),1.35(s,6H).

[0085] Referring to the conditions and steps of the second synthetic method in Example 3, naphthalene-1-ylmethyl bromide was reacted with compound 7 to obtain 3-(naphthalene-1-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-10) with a yield of 68%. MS (ESI) (m / z): 361 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.28(s,1H),8.35(d,J=7.5Hz,1H),8.21(s,1H),8.15(s,1H),8.01(s,2H ),7.91(s,1H),7.80(d,J=6.1Hz,1H),7.73(s,1H),7.61–7.44(m,4H),7.27(s,1H),5.93(s,2H).

[0086] Referring to the conditions and steps of the general synthesis method 2 in Example 3, naphthalene-2-ylmethyl bromide was reacted with compound 7 to obtain 3-(naphthalene-2-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-11) with a yield of 69%. MS (ESI) (m / z): 361 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.53(s,1H),8.61(s,1H),8.25(s,1H),8.14–7.79(m,6H),7.71(s,1H),7.52(s,2H),7.30(d,J=11.6Hz,2H),5.59(s,2H).

[0087] Referring to the conditions and steps of the general synthesis method 2 in Example 3, quinolin-8-ylmethyl bromide was reacted with compound 7 to obtain N-(pyridin-3-yl)-3-(quinolin-8-ylmethoxy)thiophene-2-carboxamide (Formula 1-12) with a yield of 78%. MS (ESI) (m / z): 362 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.57(s,1H),8.88(s,1H),8.46(d,J=8.1Hz,1H),8.27(d,J=14.2Hz,2H),8.05(d,J=7.6Hz ,2H),7.86(s,2H),7.68(d,J=7.1Hz,1H),7.57(d,J=3.7Hz,1H),7.43(s,1H),7.29(d,J=24.7Hz,1H),6.03(s,2H).

[0088] Referring to the conditions and steps of the second synthetic method in Example 3, anthracene-9-ylmethyl bromide was reacted with compound 7 to obtain 3-(anthracen-9-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-13) with a yield of 55%. MS (ESI) (m / z): 411 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.03(s,1H),8.82(s,1H),8.59(d,J=8.0Hz,2H),8.21(d,J=8.0Hz,2H),8. 10(s,1H),8.00(s,1H),7.74(s,1H),7.59(s,5H),7.36(d,J=7.2Hz,1H),7.13(s,1H),6.50(s,2H).

[0089] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-methoxynaphthalen-1-ylmethyl bromide was reacted with compound 7 to obtain 3-((2-methoxynaphthalen-1-yl)methoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-16) with a yield of 88%. MS (ESI) (m / z): 391 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.23(s,1H),8.27(d,J=8.2Hz,1H),8.19(d,J=3.9Hz,1H),8.09(d,J=9.0Hz,1H),8.02(s,1H),7.93(dd,J=16.0,6.8Hz ,2H),7.66(d,J=7.1Hz,1H),7.56(d,J=9.0Hz,1H),7.54–7.46(m,2H),7.39(d,J=7.4Hz,1H),7.26(d,J=4.0Hz,1H),5.91(s,2H),3.97(s,3H).

[0090] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 6-(2-(bromomethyl)phenoxy)hexanoic acid ethyl ester was reacted with compound 7 to prepare 6-(2-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)oxymethyl)phenoxy)hexanoic acid ethyl ester (Formula 1-18) with a yield of 32%. MS (ESI) (m / z): 469 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.43(d,J=14.0Hz,1H),8.34(d,J=8.2Hz,1H),8.25(d,J= 11.2Hz,1H),7.95(d,J=7.5Hz,1H),7.88(d,J=10.8Hz,1H),7.55(s,1H),7.46–7.3 0(m,3H),7.11(d,J=7.5Hz,1H),7.01(s,1H),5.37(s,2H),3.95(dd,J=32.5,10.5H z,4H),2.14(s,2H),1.57(s,2H),1.40(s,2H),1.25(s,2H),1.14(t,J=2.7Hz,3H).

[0091] Referring to the conditions and steps of the general synthesis method 2 in Example 3, (6,7,8,9-tetrafluorodibenzo[b,e][1,4]dioxol-1-yl)methyl bromide was reacted with compound 7 to obtain N-(pyridin-3-yl)-3-((6,7,8,9-tetrafluorodibenzo[b,e][1,4]dioxol-1-yl)methoxy)thiophene-2-carboxamide (Formula 1-19) with a yield of 45%. MS (ESI) (m / z): 489 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.41(s,1H),8.52(s,1H),8.21(s,1H),7.99(s,1H ),7.90(s,1H),7.39–7.26(m,3H),7.22(s,1H),7.17(s,1H),5.43(s,2H).

[0092] Referring to the conditions and steps of the second synthetic method in Example 3, 2-nitrobenzyl bromide was reacted with compound 7 to obtain 3-(2-nitrobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-21) with a yield of 16%. MS (ESI) (m / z): 356 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.43(s,1H),8.69(s,1H),8.44–8.02(m,3H),7.84(m,3H),7.66(s,1H),7.38(s,1H),7.24(s,1H),5.80(s,2H).

[0093] Referring to the conditions and steps of the general synthesis method 2 in Example 3, methyl 2-(bromomethyl)benzoate was reacted with compound 7 to obtain methyl 2-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)oxymethyl)benzoate (Formula 1-22) with a yield of 45%. MS (ESI) (m / z): 369 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.61(s,1H),8.27(s,1H),8.05(d,J=7.7Hz,1H),7.97(d,J=7.5Hz,1H),7.85(d,J=5.0Hz,1H) ,7.75(d,J=7.4Hz,1H),7.67(t,J=7.3Hz,1H),7.54(t,J=7.2Hz,1H),7.37(s,1H),7.25(d,J=5.3Hz,1H),5.72(s,2H),3.74(s,3H).

[0094] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-chloro-5-(trimethylacetoxy)benzyl bromide was reacted with compound 7 to prepare trimethylacetic acid (4-chloro-3-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)methoxy))phenol ester (Formula 1-23) with a yield of 36%. MS (ESI) (m / z): 445,447 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ9.43(s,1H),8.61(d,J=3.1Hz,1H),8.26(s,1H),8.00(s,1H),7.88(t,J=5.6Hz,1H ),7.58(dd,J=12.6,3.9Hz,2H),7.33(d,J=5.5Hz,2H),7.24–7.15(m,1H),5.45(s,2H),1.26–1.20(s,9H).

[0095] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2-chloro-5-(diethoxyphosphoryloxy)benzyl bromide was reacted with compound 7 to obtain 4-chloro-3-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)methoxy))phenylphosphodiethyl ester (Formula 1-25) with a yield of 85%. MS (ESI) (m / z): 497,499 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.45(s,1H),8.64(s,1H),8.29(s,1H),8.04(s,1H),7.89(s,1H),7.59(d,J=10.6Hz,2 H),7.37(s,1H),7.30(t,J=7.2Hz,2H),5.50(d,J=8.7Hz,2H),4.08(dd,J=15.2,7.5Hz,4H),1.22–1.14(m,6H).

[0096] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(dodecyloxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-(dodecyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-27) with a yield of 68%. MS (ESI) (m / z): 495 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.39 (s, 1H), 8.26 (m, 2H), 8.15 (s, 1H), 7.46 (d, J = 34.6Hz, 3H), 7.24–7 .18(m,1H),7.03(s,3H),5.31(s,2H),3.95(m,2H),1.65(m,2H),1.23(m,18H),0.88(t,3H).

[0097] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(undecanyloxy)benzyl bromide was reacted with compound 7 to prepare 3-((2-(undecanyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-28) with a yield of 52%. MS (ESI) (m / z): 481 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.39 (s, 1H), 8.28 (m, 2H), 8.15 (s, 1H), 7.47 (d, J = 33.6Hz, 3H), 7.26–7 .17(m,1H),7.04(m,3H),5.42(s,2H),3.96(m,2H),1.66(m,2H),1.22(m,16H),0.88(t,3H).

[0098] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(decyloxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-(decyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-29) with a yield of 48%. MS (ESI) (m / z): 467 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.43(d,J=18.0Hz,1H),8.31(d,J=18.7Hz,2H),8.19(d,J=18.3Hz,1H),7.60–7.40(m,3H), 7.28(d,J=5.6Hz,2H),7.08(d,J=18.7Hz,3H),5.32(s,2H),3.98(m,2H),1.68(m,2H),1.24(m,14H),0.92(t,3H).

[0099] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(nonyloxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-(nonyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-30) with a yield of 58%. MS (ESI) (m / z): 453 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.38(s,1H),8.27(m,2H),8.15(s,1H),7.50(s,1H),7.42(m,2H),7.25(s,1H),7.2 3(s,1H),7.08–6.96(m,3H),5.31(s,2H),3.95(d,J=4.9Hz,2H),1.65(m,2H),1.22(m,12H),0.86(t,3H).

[0100] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(octyloxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-(octyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-31) with a yield of 59%. MS (ESI) (m / z): 439 [M+H] + . 1H NMR(400MHz, CDCl3)δ9.38(s,1H),8.27(s,2H),8.15(s,1H),7.50(s,1H),7.42(s,2H),7.25–7.16(m,1H),7. 08–6.95(m,3H),5.31(s,2H),3.96(s,2H),1.65(d,J=5.9Hz,2H),1.35–1.13(m,10H),0.86(t,J=6.7Hz,3H).

[0101] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(heptyloxy)benzyl bromide was reacted with compound 7 to prepare 3-((2-(heptyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-32) with a yield of 52%. MS (ESI) (m / z): 425 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.38(s,1H),8.26(d,J=8.2Hz,2H),8.15(s,1H),7.51(d,J=5.0Hz,1H),7.42(m,2H),7.23(s,1H) ,7.02(dd,J=22.1,7.0Hz,3H),5.29(s,2H),3.96(t,J=6.1Hz,2H),1.70–1.61(m,2H),1.38–1.11(m,8H),0.83(t,3H).

[0102] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(hexyloxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-hexyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-33) with a yield of 56%. MS (ESI) (m / z): 411 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.39(s,1H),8.27(m,2H),8.15(s,1H),7.51(s,1H),7.42(m,2H),7. 24(m,2H),7.05(m,3H),5.32(s,2H),3.96(m,2H),1.65(m,2H),1.25(m,6H),0.81(t,3H).

[0103] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(pentyloxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-pentyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-34) with a yield of 21%. MS (ESI) (m / z): 397 [M+H] + .1 H NMR(400MHz, CDCl3)δ9.38(s,1H),8.27(m,2H),8.15(s,1H),7.51(s,1H),7.42(m,2H),7.24–7.19 (m,1H),7.02(d,J=18.8Hz,3H),5.32(s,2H),3.96(m,2H),1.65(m,2H),1.26(m,4H),0.81(t,3H).

[0104] Referring to the conditions and steps of the second synthetic method in Example 3, 2-(butoxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-butoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-35) with a yield of 30%. MS (ESI) (m / z): 383 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.38(s,1H),8.27(s,2H),8.16(s,1H),7.52(s,1H),7.43(2,2H),7.25–7.18 (m,1H),7.02(d,J=20.3Hz,3H),5.37(s,2H),3.97(m,2H),1.65(m,2H),1.35(m,2H),0.84(t,3H).

[0105] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2-(propoxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-propoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-36) with a yield of 91%. MS (ESI) (m / z): 369 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.39(s,1H),8.27(s,2H),8.17(s,1H),7.52(s,1H),7.42(m,2H),7.2 5–7.16(m,1H),7.03(d,J=22.0Hz,3H),5.33(s,2H),3.93(m,2H),1.69(m,2H),0.91(t,3H).

[0106] Referring to the conditions and steps of the second synthetic method in Example 3, 2-(ethoxy)benzyl bromide was reacted with compound 7 to obtain 3-((2-ethoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-37) with a yield of 57%. MS (ESI) (m / z): 355 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.40 (s, 1H), 8.27 (s, 2H), 8.19 (s, 1H), 7.52 (s, 1H), 7.42 (d, J = 6. 7Hz,2H),7.25–7.18(m,1H),7.11–6.90(m,3H),5.38(s,2H),4.04(m,2H),1.31(t,3H).

[0107] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2-n-pentyloxy-6-chlorobenzyl bromide was reacted with compound 7 to obtain 3-(2-n-pentyloxy-6-chlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-38) with a yield of 57%. MS (ESI) (m / z): 430, 432 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.32(s,1H),8.27(d,J=2.8Hz,2H),8.17(d,J=1.8Hz,1H),7.51(d,J=5.4Hz,1H),7.34(d,J=8.2Hz,1H),7.24(d ,J=7.8Hz,1H),7.13(dd,J=16.8,6.7Hz,2H),6.90(d,J=8.1Hz,1H),5.50(s,2H),3.97(m,2H),1.71(m,2H),1.30(m,4H),0.84(t,3H).

[0108] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-chloro-6-n-propoxybenzyl bromide was reacted with compound 7 to obtain 3-(2-chloro-6-n-propoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-39) with a yield of 52%. MS (ESI) (m / z): 403, 405 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.32(s,1H),8.28(m,2H),8.18(s,1H),7.51(s,1H),7.34(s,1H),7.25–7.20 (m,1H),7.14(d,J=15.8Hz,2H),6.90(s,1H),5.51(s,2H),3.94(m,2H),1.75(m,2H),0.94(t,3H).

[0109] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2-methylbenzyl bromide was reacted with compound 7 to obtain 3-(2-methylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-41) with a yield of 91%. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.44(s,1H),8.48(s,1H),8.28(s,1H),8.00(d,J=7.0Hz,1H),7. 89(s,1H),7.54(d,J=6.3Hz,1H),7.30(dd,J=21.5,7.0Hz,5H),5.46(s,2H),2.40(s,3H).

[0110] Referring to the conditions and steps of the second synthetic method in Example 3, 4-methylbenzyl bromide was reacted with compound 7 to obtain 3-(4-methylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-42) with a yield of 60%. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.46(s,1H),8.58(s,1H),8.29(s,1H),8.04(d,J=7.1Hz,1H),7.86(d,J=5.3Hz,1 H), 7.48 (d, J = 7.4Hz, 2H), 7.38 (d, J = 5.3Hz, 1H), 7.26 (dd, J = 15.0, 6.2Hz, 3H), 5.39 (s, 2H), 2.31 (s, 3H).

[0111] Referring to the conditions and steps of the second synthetic method in Example 3, 3-methylbenzyl bromide was reacted with compound 7 to obtain 3-(3-methylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-43) with a yield of 84%. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.49 (s, 1H), 8.58 (s, 1H), 8.28 (d, J = 4.7Hz, 1H), 8.05 (d, J = 8.3Hz, 1H), 7.87 (d, J = 5. 5Hz, 1H), 7.43 (s, 1H), 7.33 (ddd, J = 27.2, 12.9, 6.4Hz, 4H), 7.20 (d, J = 7.0Hz, 1H), 5.39 (s, 2H), 2.31 (s, 3H).

[0112] Referring to the conditions and steps of the second synthetic method in Example 3, 2-fluorobenzyl bromide was reacted with compound 7 to obtain 3-(2-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-44) with a yield of 67%. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.42 (s, 1H), 8.57 (s, 1H), 8.29 (d, J = 4.4Hz, 1H), 8.03 (d, J = 8.0Hz, 1H), 7.89(d,J=5.5Hz,1H),7.70(t,J=7.4Hz,1H),7.58–7.43(m,1H),7.44–7.23(m,4H),5.52(s,2H).

[0113] Referring to the conditions and steps of the second synthetic method in Example 3, 3-fluorobenzyl bromide was reacted with compound 7 to obtain 3-(3-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-45) with a yield of 54%. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.34(s,1H),8.50(s,1H),8.15(s,1H),7.94(s,1H),7.72(s,1H),7.31(d,J=22.9Hz,4H),7.10(m,2H),5.31(d,J=7.5Hz,2H).

[0114] Referring to the conditions and steps of the second synthetic method in Example 3, 4-fluorobenzyl bromide was reacted with compound 7 to obtain 3-(4-fluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-46) with a yield of 41%. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.45 (s, 1H), 8.60 (s, 1H), 8.29 (s, 1H), 8.04 (d, J = 7.4Hz, 1H), 7.8 6(d,J=5.2Hz,1H),7.67(m,2H),7.38(d,J=4.7Hz,1H),7.27(t,J=7.5Hz,3H),5.42(s,2H).

[0115] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2-chlorobenzyl bromide was reacted with compound 7 to obtain 3-(2-chlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-47) with a yield of 48%. MS (ESI) (m / z): 345, 347 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ9.42(s,1H),8.55(s,1H),8.29(d,J=4.1Hz,1H),8.03(d,J=8.1Hz,1H),7. 90(d,J=5.5Hz,1H),7.74(d,J=6.8Hz,1H),7.58(d,J=7.3Hz,1H),7.51–7.30(m,4H),5.52(s,2H).

[0116] Referring to the conditions and steps of the second synthetic method in Example 3, 3-chlorobenzyl bromide was reacted with compound 7 to obtain 3-(3-chlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-48) with a yield of 56%. MS (ESI) (m / z): 345, 347 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.49(s,1H),8.67(s,1H),8.29(s,1H),8.07(d,J=8.2Hz,1H),7.87(d,J=5.5Hz, 1H),7.73(s,1H),7.54(s,1H),7.51–7.42(m,2H),7.41–7.32(m,1H),7.25(d,J=5.3Hz,1H),5.44(s,2H).

[0117] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 4-chlorobenzyl bromide was reacted with compound 7 to obtain 3-(4-chlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-49) with a yield of 63%. MS (ESI) (m / z): 345, 347 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.46 (s, 1H), 8.63 (s, 1H), 8.29 (d, J = 4.2Hz, 1H), 8.06 (d, J = 8.3Hz, 1H), 7.86 (d, J = 5. 5Hz,1H),7.62(d,J=8.1Hz,2H),7.50(d,J=8.1Hz,2H),7.44–7.35(m,1H),7.25(d,J=5.5Hz,1H),5.43(s,2H).

[0118] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-cyanobenzyl bromide was reacted with compound 7 to obtain 3-(2-cyanobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-50) with a yield of 51%. MS (ESI) (m / z): 336 [M+H] +. 1 H NMR (400MHz, CDCl3) δ9.01 (s, 1H), 8.32 (d, J = 13.9Hz, 3H), 7.65 (m, 5H), 7.25 (m, 1H), 7.00 (s, 1H), 5.50 (s, 2H).

[0119] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 3-cyanobenzyl bromide was reacted with compound 7 to obtain 3-(3-cyanobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-51) with a yield of 54%. MS (ESI) (m / z): 336 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.01(s,1H),8.31(m,2H),8.23(s,1H),7.77(m,3H),7.63(s,1H),7.53(s,1H),6.93(s,1H),5.34(s,2H).

[0120] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 4-cyanobenzyl bromide was reacted with compound 7 to obtain 3-(4-cyanobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-52) with a yield of 46%. MS (ESI) (m / z): 336 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.02(s,1H),8.34(d,J=14.2Hz,2H),8.23(s,1H),7.78(d,J=8.0Hz,2H), 7.61 (d, J = 7.3Hz, 2H), 7.52 (d, J = 5.4Hz, 1H), 7.27 (s, 1H), 6.90 (d, J = 5.5Hz, 1H), 5.38 (s, 2H).

[0121] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-trifluoromethoxybenzyl bromide was reacted with compound 7 to obtain 3-(2-trifluoromethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-53) with a yield of 51%. MS (ESI) (m / z): 395 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.09 (s, 1H), 8.27 (m, 1H), 7.60 (d, J = 7.3Hz, 3H), 7.52 (s, 1H), 7.41 (m, 2H), 7.29 (m, 2H), 7.00 (s, 1H), 5.39 (s, 2H).

[0122] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 4-trifluoromethoxybenzyl bromide was reacted with compound 7 to obtain 3-(4-trifluoromethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-54) with a yield of 70%. MS (ESI) (m / z): 395 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.12(s,1H),8.41(s,1H),8.32(s,1H),8.13(s,1H),7.53(m,3H),7.26(m,3H),6.96(s,1H),5.30(s,2H).

[0123] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 3-trifluoromethoxybenzyl bromide was reacted with compound 7 to obtain 3-(3-trifluoromethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-55) with a yield of 55%. MS (ESI) (m / z): 395 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.10 (s, 1H), 8.32 (m, 2H), 8.18 (s, 1H), 7.49 (d, J = 29.7Hz, 3H), 7.34 (m, 2H), 7.23 (s, 1H), 6.96 (s, 1H), 5.31 (s, 2H).

[0124] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 4-methoxybenzyl bromide was reacted with compound 7 to obtain 3-(4-methoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-56) with a yield of 23%. MS (ESI) (m / z): 341 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.24 (s, 1H), 8.26 (d, J = 17.2Hz, 3H), 7.51 (s, 1H), 7.41 (s, 2H), 7.20 (m, 1H), 6.99 (s, 3H), 5.21 (s, 2H), 3.85 (s, 3H).

[0125] Referring to the conditions and steps of the second synthetic method in Example 3, 2-methoxybenzyl bromide was reacted with compound 7 to obtain 3-(2-methoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-57) with a yield of 78%. MS (ESI) (m / z): 341 [M+H] + . 1H NMR(400MHz, CDCl3)δ9.39(s,1H),8.28(s,1H),8.22(m,2H),7.51(s,1H),7.4 2(s,1H),7.23(m,2H),7.02(dd,J=20.4,6.7Hz,3H),5.32(s,2H),3.82(s,3H).

[0126] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 3-methoxybenzyl bromide was reacted with compound 7 to obtain 3-(3-methoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-58) with a yield of 46%. MS (ESI) (m / z): 341 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.23(s,1H),8.29(m,3H),7.45(m,2H),7.29(s,1H),7.00(m,4H),5.26(s,2H),3.83(s,3H).

[0127] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-trifluoromethylbenzyl bromide was reacted with compound 7 to obtain 3-(2-trifluoromethylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-59) with a yield of 48%. MS (ESI) (m / z): 379 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.09 (s, 1H), 8.26 (m, 3H), 7.66 (dd, J = 81.3, 36.6Hz, 5H), 7.18 (m, 1H), 6.97 (s, 1H), 5.51 (s, 2H).

[0128] Referring to the conditions and steps of the second synthetic method in Example 3, 2,6-dimethylbenzyl bromide was reacted with compound 7 to obtain 3-(2,6-dimethylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-60) with a yield of 55%. MS (ESI) (m / z): 339 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.19(s,1H),8.24(s,1H),8.11(s,1H),7.91(s,1H),7. 56(s,1H),7.28(s,1H),7.18(m,3H),7.10(s,1H),5.34(s,2H),2.46(s,6H).

[0129] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2-chloro-6-methoxybenzyl bromide was reacted with compound 7 to obtain 3-(2-chloro-6-methoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-63) with a yield of 45%. MS (ESI) (m / z): 375, 377 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.37(s,1H),8.32(m,2H),8.26(s,1H),7.55(s,1H),7.41(s,1H) ,7.27(m,1H),7.18(d,J=8.0Hz,2H),6.96(d,J=7.9Hz,1H),5.55(s,2H),3.90(s,3H).

[0130] Referring to the conditions and steps of the general synthetic method 2 in Example 3, benzo[b]thiophen-3-ylmethyl bromide was reacted with compound 7 to obtain 3-(benzo[b]thiophen-3-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-66) with a yield of 34%. MS (ESI) (m / z): 367 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.99(s,1H),8.20(s,1H),7.98(s,1H),7.88(d,J=8.8Hz,2H ),7.76(s,1H),7.63(s,1H),7.56(s,1H),7.40(m,2H),7.09(m,2H),5.53(s,2H).

[0131] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2,3,5,6-tetrafluorobenzyl bromide was reacted with compound 7 to obtain 3-(2,3,5,6-tetrafluorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-67) with a yield of 65%. MS (ESI) (m / z): 383 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.98(s,1H),8.49(s,1H),8.35(d,J=3.6Hz,1H),8.26(d,J=8.2Hz,1H),7.5 4(d,J=5.5Hz,1H),7.29(d,J=8.0Hz,1H),7.23–7.14(m,1H),7.07(d,J=5.5Hz,1H),5.49(s,2H).

[0132] Referring to the conditions and steps of the second synthetic method in Example 3, 2-(phenethoxy)benzyl bromide was reacted with compound 7 to obtain 3-(2-(phenethoxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-70) with a yield of 80%. MS (ESI) (m / z): 431 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.30(s,1H),8.27(m,2H),8.11(s,1H),7.52(s,1H),7.40(m,2H),7 .25–7.19(m,1H),7.05(dd,J=45.4,17.6Hz,8H),5.25(s,2H),4.20(t,2H),2.96(t,2H).

[0133] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 4-chloro-2-hexyloxybenzyl bromide was reacted with compound 7 to obtain 3-(4-chloro-2-hexyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-71) with a yield of 60%. MS (ESI) (m / z): 445, 447 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.27(s,1H),8.28(d,J=12.0Hz,3H),7.51(s,1H),7.45–7.32(m,2H),7.26–7.17(m, 1H), 7.00 (s, 1H), 6.92 (d, J = 8.6Hz, 1H), 5.26 (s, 2H), 3.94 (m, 2H), 1.66 (m, 2H), 1.25 (m, 6H), 0.82 (t, 3H).

[0134] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 4-chloro-2-heptyloxybenzyl bromide was reacted with compound 7 to obtain 3-(4-chloro-2-heptyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-72) with a yield of 70%. MS (ESI) (m / z): 459, 461 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.25(s,1H),8.26(d,J=7.6Hz,3H),7.51(s,1H),7.38(d,J=15.2Hz,2H),7.25(s,2 H), 7.00 (s, 1H), 6.90 (d, J = 7.7Hz, 1H), 5.26 (s, 2H), 3.93 (m, 2H), 1.65 (m, 2H), 1.23 (m, 8H), 0.83 (t, 3H).

[0135] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 5-chloro-2-hexyloxybenzyl bromide was reacted with compound 7 to obtain 3-(5-chloro-2-hexyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-73) with a yield of 15%. MS (ESI) (m / z): 445, 447 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.27 (s, 1H), 8.28 (d, J = 10.5Hz 3H),7.52(s,1H),7.40(d,J=14.8Hz,3H),7.01(s,1H),6.93(s,1H),5.26(s,2H),3.95(m,2H),1.66(m,2H),1.26(m,6H),0.82(t,3H).

[0136] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 5-chloro-2-heptyloxybenzyl bromide was reacted with compound 7 to obtain 3-(5-chloro-2-heptyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-74) with a yield of 56%. MS (ESI) (m / z): 459, 461 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.07(s,1H),8.07(m,3H),7.32(s,1H),7.22(m,2H),7.08(s,1H),6. 81(s,1H),6.73(s,1H),5.07(s,2H),3.74(m,2H),1.47(m,2H),1.05(m,8H),0.65(t,3H).

[0137] Referring to the conditions and steps of the second synthetic method in Example 3, 2-(4-chlorobutyloxy)benzyl bromide was reacted with compound 7 to obtain 3-(2-(4-chlorobutyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-75) with a yield of 48%. MS (ESI) (m / z): 417, 419 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.43(s,1H),8.34(s,1H),8.19(s,1H),7.60(s,1H),7.50–7.41(m,1H),7 .38–7.19(m,2H),7.09(d,J=30.2Hz,2H),5.49(s,2H),4.08(m,2H),3.58(m,2H),1.90(m,4H).

[0138] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(6-chlorohexyloxy)benzyl bromide was reacted with compound 7 to obtain 3-(2-(6-chlorohexyloxy)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-76) with a yield of 47%. MS (ESI) (m / z): 445, 447 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.40 (d, J = 47.7Hz, 1H), 8.27 (d, J = 57.3Hz, 3H), 7.60 (s, 1H), 7.32 (d, J = 16.6 Hz, 2H), 7.09 (d, J = 23.1Hz, 3H), 5.46 (s, 2H), 4.04 (m, 2H), 3.46 (m, 2H), 1.73 (m, 4H), 1.42 (m, 4H).

[0139] Referring to the conditions and steps of the second synthetic method in Example 3, thiophene-2-ylmethyl bromide was reacted with compound 7 to obtain 3-(thiophene-2-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-77) with a yield of 9%. MS (ESI) (m / z): 317 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.18 (s, 1H), 8.32 (d, J = 19.4Hz, 3H), 7.51 (d, J = 27.6Hz, 2H), 7.26–7.16 (m, 2H), 7.06 (d, J = 26.8Hz, 2H), 5.45 (s, 2H).

[0140] Referring to the conditions and steps of the second synthetic method in Example 3, 3-(thiophen-3-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-78) was prepared by reacting thiophen-3-ylmethyl bromide with compound 7 with a yield of 8%. MS (ESI) (m / z): 317 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.21(s,1H),8.30(m,3H),7.60–7.42(m,3H),7.30(m,2H),6.99(s,1H),5.30(s,2H).

[0141] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 3-hexyloxybenzyl bromide was reacted with compound 7 to obtain 3-(3-hexyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-79) with a yield of 12%. MS (ESI) (m / z): 411 [M+H] + . 1H NMR(400MHz, CDCl3)δ9.24(s,1H),8.31(m,2H),8.20(s,1H),7.51(s,1H),7.38(s,1H),7.27(s,1H),7.23(s,1H), 7.06(s,1H),6.98(d,J=8.1Hz,3H),5.24(s,2H),3.95(m,2H),1.76(m,2H),1.43(m,2H),1.32(m,4H),0.89(t,3H).

[0142] Referring to the conditions and steps of the second synthetic method in Example 3, 3-chloro-2-hexyloxybenzyl bromide was reacted with compound 7 to obtain 3-(3-chloro-2-hexyloxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-80) with a yield of 54%. MS (ESI) (m / z): 445, 447 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.20(s,1H),8.41(s,1H),8.27(d,J=28.6Hz,2H),7.50(d,J=14.1Hz,2H),7.37(s,1H ),7.16(m,2H),7.02(s,1H),5.31(s,2H),4.04(m,2H),1.75(m,2H),1.38(m,2H),1.22(m,4H),0.82(t,3H).

[0143] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-ethylbenzyl bromide was reacted with compound 7 to obtain 3-(2-ethylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-81) with a yield of 82%. MS (ESI) (m / z): 339 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.20 (s, 1H), 8.26 (d, J = 4.3Hz, 1H), 8.17 (d, J = 7.8Hz, 1H), 8.07 (s, 1H), 7.54 (d, J = 4.5Hz, 1H), 7.44 (t, J = 7.8Hz, 2H),7.34(dd,J=16.6,7.1Hz,2H),7.24–7.18(m,1H),7.04(d,J=4.4Hz,1H),5.31(s,2H),2.79(q,J=7.2Hz,2H),1.26(t,J=7.2Hz,3H).

[0144] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-isopropoxybenzyl bromide was reacted with compound 7 to obtain 3-(2-isopropoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-82) with a yield of 43%. MS (ESI) (m / z): 369 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.37 (s, 1H), 8.37–8.17 (m, 3H), 7.52 (d, J = 5.5Hz, 1H), 7.43 (dd, J = 15.9, 6.9Hz, 2H), 7.2 3(dd,J=8.2,4.9Hz,1H),7.07–6.97(m,2H),5.32(s,2H),4.61(dt,J=12.0,5.9Hz,1H),1.27(d,J=6.0Hz,6H).

[0145] Referring to the conditions and steps of the general synthetic method 2 in Example 3, ethyl 2-bromomethylbenzoate was reacted with compound 7 to prepare ethyl 2-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)oxymethyl)benzoate (Formula 1-84) with a yield of 53%. MS (ESI) (m / z): 383 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.94 (s, 1H), 8.24 (dd, J = 7.5, 1.3Hz, 1H), 8.08 (dt, J = 7.5 ,1.4Hz,1H),8.02(d,J=1.3Hz,1H),7.89(d,J=7.5Hz,1H),7.77(dd,J=7.4,1.4H z,1H),7.64(td,J=7.5,1.4Hz,1H),7.56–7.46(m,2H),7.26(t,J=7.5Hz,1H),6 .55(d,J=7.5Hz,1H),5.20(s,2H),4.31(q,J=6.0Hz,2H),1.38(t,J=6.0Hz,3H).

[0146] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-difluoromethoxybenzyl bromide was reacted with compound 7 to obtain 3-(2-difluoromethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-85) with a yield of 47%. MS (ESI) (m / z): 377 [M+H] + . 1H NMR (400MHz, CDCl3) δ9.16 (s, 1H), 8.27 (m, 3H), 7.53 (d, J = 15.4Hz, 3H), 7.34 (s,1H),7.25–7.16(m,2H),7.01(s,1H),6.60(t,J=73.2Hz,1H),5.37(s,2H).

[0147] Referring to the conditions and steps of the second synthetic method in Example 3, 2-bromobenzyl bromide was reacted with compound 7 to obtain 3-(2-bromobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-86) with a yield of 36%. MS (ESI) (m / z): 389, 391 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.17 (s, 1H), 8.30 (m, 3H), 7.71 (d, J = 7.8Hz, 1H), 7.52 (m, 2H), 7.42 (s, 1H), 7.33 (s, 2H), 7.01 (s, 1H), 5.38 (s, 2H).

[0148] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2-iodobenzyl bromide was reacted with compound 7 to obtain 3-(oxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-87) with a yield of 39%. MS (ESI) (m / z): 436,438 [M+H] + . 1 HNMR (400MHz, CDCl3) δ9.14(s,1H),8.28(m,3H),7.98(s,1H),7.57–7.41(m,3H),7.25–7.21(m,1H),7.17(s,1H),7.00(s,1H),5.32(s,2H).

[0149] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 5-fluoro-2-methylbenzyl bromide was reacted with compound 7 to obtain 3-(5-fluoro-2-methylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-88) with a yield of 38%. MS (ESI) (m / z): 343 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.11(s,1H),8.30(s,1H),8.21(m,2H),7.54(s,1H),7.28(s,1H),7.2 6–7.21(m,1H),7.16(d,J=8.4Hz,1H),7.07(s,1H),6.99(s,1H),5.26(s,2H),2.40(s,3H).

[0150] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2,5-dichlorobenzyl bromide was reacted with compound 7 to obtain 3-(2,5-dichlorobenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-89) with a yield of 48%. MS (ESI) (m / z): 379,381 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.07 (s, 1H), 8.37 (d, J = 33.1Hz, 3H), 7.54 (m, 2H), 7.42 (d, J = 27.6Hz, 2H), 6.98 (s, 1H), 5.37 (s, 2H).

[0151] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2,5-dipropoxybenzyl bromide was reacted with compound 7 to obtain 3-(2,5-dipropoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-93) with a yield of 90%. MS (ESI) (m / z): 427 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.36 (s, 1H), 8.25 (d, J = 14.6Hz, 3H), 7.53 (d, J = 29.3Hz, 1H), 7.24 (s, 1H), 7 .11–6.86(m,4H),5.29(s,2H),3.87(m,4H),1.77(m,2H),1.68(m,2H),1.01(t,3H),0.90(t,3H).

[0152] Referring to the conditions and steps of the general synthesis method 2 in Example 3, (1,1'-biphenyl-2-yl)methyl bromide was reacted with compound 7 to obtain 3-((1,1'-biphenyl-2-yl)methoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-94) with a yield of 41%. MS (ESI) (m / z): 387 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.29(s,1H),8.45(s,1H),8.28(s,1H),7.96(s,1H),7.79(m,2H),7.49(m,2H),7.39(m,7H),6.97(s,1H),5.36(s,2H).

[0153] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 3-trifluoromethylbenzyl bromide was reacted with compound 7 to obtain 3-(3-trifluoromethylbenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-97) with a yield of 43%. MS (ESI) (m / z): 379 [M+H]+ . 1 H NMR(400MHz,DMSO-d6)δ9.50(s,1H),8.65(s,1H),8.28(s,1H),8.03(m,2H),7.88(d,J=4.2Hz,2H),7.75 (d, J=7.1Hz, 1H), 7.69 (d, J=7.3Hz, 1H), 7.37 (dd, J=7.9, 4.4Hz, 1H), 7.29 (d, J=4.9Hz, 1H), 5.52 (s, 2H).

[0154] Referring to the conditions and steps of the general synthesis method 2 in Example 3, 2,5-dimethoxybenzyl bromide was reacted with compound 7 to obtain 3-(2,5-dimethoxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-98) with a yield of 52%. MS (ESI) (m / z): 371 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.51(s,1H),8.54(s,1H),8.29(s,1H),8.01(d,J=8.5Hz,1H),7.87(d,J=5.5Hz,1H ),7.46–7.32(m,2H),7.17(s,1H),7.03(d,J=9.1Hz,1H),6.96(d,J=8.9Hz,1H),5.38(s,2H),3.71(s,6H).

[0155] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 3-(adamantan-1-ylmethoxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-99) was prepared by reacting adamantane-1-ylmethyl bromide with compound 7 in a yield of 98%. MS (ESI) (m / z): 369 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.29(s,1H),8.72(s,1H),8.31(s,1H),8.15(s,1H),7.86(d,J=5.3 Hz,1H),7.52–7.38(m,1H),7.21(d,J=5.4Hz,1H),3.91(s,2H),2.00(m,3H),1.67(m,12H).

[0156] Example 5: Preparation of 3-benzyloxy-N-(2-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 1-90)

[0157] 1) Synthesis of 3-benzyloxythiophene-2-carbonyl chloride

[0158]

[0159] Weigh 1g (4.27mmol, 1.0eq) of compound 5 into a round-bottom flask, add 50mL DCM to dissolve, add 5.4g (42.7mmol, 10.0eq) of oxalyl chloride while stirring in an ice bath, then add 500μLDMF as catalyst, stir at room temperature for 7h, TLC detection shows that the reaction is complete, evaporate to dryness, add EA, wash once with saturated brine, dry over anhydrous sodium sulfate, evaporate to dryness to obtain 996mg of a light yellow solid, with a yield of 92%. The product can be used in the next reaction without further purification.

[0160] 2) Synthesis of 3-benzyloxy-N-(2-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 1-90) (General Synthesis Method 3)

[0161]

[0162] 71 mg (0.64 mmol, 2.0 eq) of compound 10 was weighed into a round-bottom flask, 4 mL of DCM was added to dissolve, 82 mg (0.64 mmol, 2.0 eq) of DIPEA was added, and 80 mg (0.32 mmol, 1.0 eq) of compound 9 was added under stirring, and stirred at room temperature overnight. The reaction was complete when detected by TLC, and the reaction was stopped. After flash purification, 100 mg of white solid was obtained, namely 3-(benzyloxy)-N-(2-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 1-90), with a yield of 96%. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.45(s,1H),8.89(s,1H),7.82(s,1H),7.53–7.39(m,5H),7.26–7.23(m,1H),7.16(s,1H),6.98(s,1H),5.31(s,2H).

[0163] Example 6: Preparation of Compounds 1-62, 1-91, 1-92

[0164] Referring to the conditions and steps of the general method 3 in Example 5, 3-methylaminopyridine was reacted with compound 9 to obtain 3-benzyloxy-N-methyl-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-62) with a yield of 27%. MS (ESI) (m / z): 325 [M+H] + . 1H NMR (400MHz, CDCl3) δ8.35(d,J=7.5Hz,2H),7.34(t,J=11.0Hz,4H),7.23(d,J=5.4Hz,1 H), 7.19 (d, J = 6.6Hz, 2H), 7.07 (s, 1H), 6.50 (d, J = 4.2Hz, 1H), 4.73 (s, 2H), 3.44 (s, 3H).

[0165] Referring to the conditions and steps of the general method 3 in Example 5, 3-amino-4-chloropyridine was reacted with compound 9 to obtain 3-benzyloxy-N-(4-chloropyridin-3-yl)thiophene-2-carboxamide (Formula 1-91) with a yield of 26%. MS (ESI) (m / z): 345, 347 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.79(s,1H),9.51(s,1H),8.20(s,1H),7.54–7.40(m,5H),7.26(s,1H),7.20(s,1H),6.99(s,1H),5.28(s,2H).

[0166] Referring to the conditions and steps of the general method 3 in Example 5, 3-amino-5-fluoropyridine was reacted with compound 9 to obtain 3-benzyloxy-N-(5-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 1-92) with a yield of 98%. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.36 (s, 1H), 8.22 (d, J = 9.7Hz, 1H), 8.16 (s, 1H), 7.89 (s ,1H),7.56(d,J=5.2Hz,1H),7.51(m,5H),7.02(d,J=4.8Hz,1H),5.30(s,2H).

[0167] Example 7: Preparation of Compounds 1-95 and 1-96

[0168]

[0169] 50 mg (0.23 mmol, 1.0 eq) of compound 7 and 63 mg (0.45 mmol, 2.0 eq) of potassium carbonate were weighed into a round-bottom flask, 3 mL of NMP was added to dissolve, 50 mg of compound 11 was added under stirring, and the mixture was moved into a 70°C oil bath and stirred overnight. TLC detected that the reaction was complete, EA was added to dissolve, the mixture was washed with saturated brine 3 times, dried over anhydrous sodium sulfate, and flash purified to obtain 30 mg of a yellow solid with a yield of 48%. MS (ESI) (m / z): 277 [M+H]+ . 1 H NMR(400MHz,DMSO-d6)δ9.40(s,1H),8.75(s,1H),8.31(s,1H),8.12(s,1H),7.87(s ,1H),7.42(s,1H),7.22(s,1H),4.33(m,2H),1.85(m,2H),1.49(m,2H),0.97(t,3H).

[0170] Referring to the preparation conditions and steps of compound 1-95, 8-iodooctane was reacted with compound 7 to obtain 3-octyloxy-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-96) with a yield of 54%. MS (ESI) (m / z): 333 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.39(s,1H),8.74(s,1H),8.30(s,1H),8.10(s,1H),7.86(s,1H ),7.39(s,1H),7.21(s,1H),4.30(m,2H),1.84(m,2H),1.49–1.18(m,10H),0.80(t,3H).

[0171] Example 8: Preparation of Compounds 1-17 and 1-24

[0172]

[0173] Weigh 100 mg of compound 1-18 into a 100 mL round-bottom flask and add a hydroxide solution (2 mol / L, H 2 O / dioxane=1 / 1)20mL, transferred to oil bath and refluxed for 8h. TLC detected that the reaction was complete, cooled, adjusted pH to neutral with dilute hydrochloric acid, extracted with EA 3 times, combined EA phases, washed once with saturated brine, dried over anhydrous sodium sulfate, and flash purified to obtain 80mg of white solid, i.e. 6-(2-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)oxymethyl)phenoxy)hexanoic acid (Formula 1-17), yield 85%. MS(ESI)(m / z): 441[M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.45 (s, 1H), 8.36 (s, 1H), 8.26 (d, J = 4.4Hz, 1H), 7.95 (d, J = 7 .5Hz,1H),7.87(d,J=5.4Hz,1H),7.53(d,J=7.3Hz,1H),7.43–7.28(m,3H),7.08(d,J =8.2Hz,1H),7.00(t,J=7.3Hz,1H),5.37(s,2H),3.92(t,J=6.0Hz,2H),2.07(t,J=7. 3Hz, 2H), 1.55 (d, J = 6.4Hz, 2H), 1.37 (dd, J = 14.4, 7.2Hz, 2H), 1.25 (d, J = 6.1Hz, 2H).

[0174] Referring to the preparation conditions and steps of compound 1-17, 3-(2-chloro-5-hydroxybenzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 1-24) was prepared using compound 1-23 as a reactant with a yield of 69%. MS (ESI) (m / z): 361, 363 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.88(s,1H),9.43(s,1H),8.59(s,1H),8.29(d,J=4.4Hz,1H),8.05(d,J=8.2Hz,1H),7.89(d,J=5.4Hz, 1H), 7.38 (dd, J=8.0, 4.8Hz, 1H), 7.33 (d, J=8.6Hz, 1H), 7.28 (d, J=5.4Hz, 1H), 7.08 (s, 1H), 6.81 (d, J=8.6Hz, 1H), 5.43 (s, 2H).

[0175] Example 9: Phosphoric acid (4-chloro-3-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)methoxy))phenol ester (Formula 1-26)

[0176]

[0177] 373 mg (0.75 mmol, 1.0 eq) of compound 1-25 was weighed into a round-bottom flask, 12 mL of acetonitrile was added to dissolve, 1.15 g (7.5 mmol, 10.0 eq) of TMS-Br was added under stirring, and the mixture was stirred at room temperature for 48 h. A large amount of white solid was precipitated. Acetonitrile was added and ultrasonic filtration was performed to obtain 246 mg of white solid, namely (4-chloro-3-((2-(pyridin-3-ylcarbamoyl)thiophen-3-yl)methoxy))phenol phosphate (Formula 1-26), with a yield of 74%. MS (ESI) (m / z): 441,443 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ9.87(s,1H),9.14(s,1H),8.57(s,1H),8.40(d,J=8.2Hz,1H),7.95(d,J=5.3Hz,1H),7.8 7(d,J=4.6Hz,1H),7.53(d,J=8.6Hz,1H),7.49(s,1H),7.32(d,J=5.2Hz,1H),7.24(d,J=8.5Hz,1H),5.50(s,2H).

[0178] Example 10: Preparation of the hydrochloride salt of compound formula 1-2

[0179] 0.60 g (1.66 mmol, 1.0 eq) of compound 1-2 was dissolved in 10 mL of dry ethyl acetate. 1.44 mL (1.8 mmol, 1.1 eq) of HCl (g) in ethyl acetate (c = 1.25 mol / L) was added dropwise to the above solution under ice-water bath conditions. After reacting for 10 min, the mixture was filtered and dried to obtain 0.53 g of a white powdery solid with a yield of 81%.

[0180] Example 11: Preparation of Compound Formula 1-100

[0181] Referring to the conditions and steps of the first synthetic method in Example 1, 4-(benzyloxy)thiophene-3-carboxylic acid and 3-aminopyridine were used as raw materials to obtain 4-(benzyloxy)-N-(pyridin-3-yl)thiophene-3-carboxamide (Formula 1-100) by amide condensation, with a yield of 80%. MS (ESI) (m / z): 311 [M+H] + . 1 H NMR(400MHz,dmso)δ9.89(s,1H),8.63(s,1H),8.27(s,1H),8.17(s,1H),8.07(d, J=8.0Hz,1H),7.57(d,J=7.2Hz,2H),7.46–7.33(m,4H),6.94(s,1H),5.21(s,2H)

[0182] Comparative Example: Preparation of Compounds 12-1 to 12-20

[0183] Referring to the conditions and steps of the first synthetic method in Example 1, 3-(2-ethylbenzyloxy)thiophene-2-carboxylic acid was reacted with compound 6 to obtain 2-(2-ethylbenzyloxy)-N-(pyridin-3-yl)benzamide (Formula 12-1) with a yield of 72%. MS (ESI) (m / z): 333 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ11.06(s,1H),9.18(d,J=1.6Hz,1H),8.60(d,J=5.3Hz,1H), 8.44(d,J=8.5Hz,1H),7.94(dd,J=8.5,5.4Hz,1H),7.64(dd,J=7.6,1.6Hz,1H),7.6 1–7.55(m,1H),7.49(d,J=7.4Hz,1H),7.40(d,J=8.3Hz,1H),7.26(dt,J=13.9,6.5H z, 2H), 7.19–7.10 (m, 2H), 5.27 (s, 2H), 2.68 (q, J = 7.5Hz, 2H), 1.12 (t, J = 7.5Hz, 3H).

[0184] Referring to the conditions and steps of the first synthetic method in Example 1, 4-aminopyridazine was reacted with compound 5 to obtain 3-benzyloxy-N-(pyridazin-4-yl)thiophene-2-carboxamide (Formula 12-2) with a yield of 96%. MS (ESI) (m / z): 312 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.74(s,1H),9.19(d,J=1.8Hz,1H),9.05(d,J=5.9Hz,1H),7.97(dd,J=5.9,2.7Hz,1H),7 .94(d,J=5.5Hz,1H),7.60(s,1H),7.59(s,1H),7.41(dt,J=22.3,7.2Hz,3H),7.29(d,J=5.5Hz,1H),5.46(s,2H).

[0185] Referring to the conditions and steps of the first synthetic method in Example 1, 2-amino-1,3,4-thiadiazole was reacted with compound 5 to obtain 3-benzyloxy-N-(1,3,4-thiadiazole-2-yl)thiophene-2-carboxamide (Formula 12-3) with a yield of 33%. MS (ESI) (m / z): 318 [M+H] + . 1 H NMR (400MHz, CDCl3) δ10.65(s,1H),8.84(s,1H),7.57(s,1H),7.45(m,4H),7.27(s,1H),6.92(s,1H),5.41(s,2H).

[0186] Referring to the conditions and steps of the first synthetic method in Example 1, 2-amino-4-fluoropyridine was reacted with compound 5 to obtain 3-benzyloxy-N-(4-fluoropyridin-2-yl)thiophene-2-carboxamide (Formula 12-4) with a yield of 38%. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.89 (s, 1H), 8.23 ​​(s, 1H), 8.13 (d, J = 9.1Hz, 1H), 7.45 (d, J = 21.3Hz, 3H), 7.27 (s, 1H), 6.89 (s, 1H), 6.77 (s, 1H), 5.39 (s, 2H).

[0187] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(1,3-dioxoisoindol-2-yl)benzyl bromide was reacted with compound 7 to prepare 3-(2-(1,3-dioxoisoindol-2-yl)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 12-5) with a yield of 50%. MS (ESI) (m / z): 456 [M+H] + . 1 H NMR(400MHz, CDCl3)δ9.97(s,1H),8.44(m,1H),8.34(m,1H),8.13(s,1H),7.87(m,1H),7.79–7.72 (m,4H),7.61(s,1H),7.49–7.37(m,2H),7.32(t,1H),7.24–7.19(m,1H),6.63(s,1H),5.24(s,2H).

[0188] Referring to the conditions and steps of the first synthetic method in Example 1, 3-aminodibenzo[b,d]furan was reacted with compound 5 to obtain 3-(benzyloxy)-N-(dibenzo[b,d]furan-3-yl)thiophene-2-carboxamide (Formula 12-6) with a yield of 75%. MS (ESI) (m / z): 400 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.65(s,1H),8.12(s,1H),8.06(t,J=6.9Hz,2H),7.88(d,J=5.4Hz,1H),7.66( dd,J=17.7,7.5Hz,3H),7.51–7.35(m,5H),7.31(d,J=5.2Hz,1H),7.24(d,J=8.2Hz,1H),5.46(s,2H).

[0189] Referring to the conditions and steps of the first synthetic method in Example 1, 2-amino-9H-fluorene was reacted with compound 5 to obtain 3-(benzyloxy)-N-(9H-fluorene-2-yl)thiophene-2-carboxamide (Formula 12-7) with a yield of 38%. MS (ESI) (m / z): 398 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ9.48(s,1H),7.83(dd,J=11.6,6.7Hz,3H),7.77(s,1H),7.62(d,J=7.0 Hz, 2H), 7.56 (d, J = 7.0 Hz, 1H), 7.52–7.39 (m, 4H), 7.39–7.23 (m, 3H), 5.45 (s, 2H), 3.90 (s, 2H).

[0190] Referring to the conditions and steps of the general synthetic method 2 in Example 3, 2-(2-((5-methylpyridin-2-yl)amino)-2-oxoethyl)benzyl bromide was reacted with compound 7 to prepare 3-(2-(2-((5-methylpyridin-2-yl)amino)-2-oxoethyl)benzyloxy)-N-(pyridin-3-yl)thiophene-2-carboxamide (Formula 12-8) with a yield of 63%. MS (ESI) (m / z): 459 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.66(s,1H),9.26(s,1H),8.42(s,1H),8.24(s,1H),8.05(s,1H),7.93(d,J=8.3Hz,1H),7.84(s,1H) ,7.74(d,J=8.7Hz,1H),7.58(s,1H),7.47–7.38(m,3H),7.31(dd,J=21.7,5.8Hz,3H),5.48(s,2H),3.95(s,2H),2.19(s,3H).

[0191] Referring to the conditions and steps of the first synthetic method in Example 1, 4-(benzyloxy)-2-methyl-N-(pyridin-3-yl)thiazole-5-carboxamide (Formula 12-9) was prepared by reacting 4-(benzyloxy)-2-methyl-N-(pyridin-3-yl)thiazole-5-carboxamide with compound 6 in a yield of 77%. MS (ESI) (m / z): 326 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ9.19(s,1H),8.46(s,1H),8.12(s,1H),7.85(d,J=8.0Hz,1 H), 7.41 (d, J = 6.9 Hz, 2H), 7.23 (dd, J = 14.6, 6.8 Hz, 4H), 5.41 (s, 2H), 2.50 (s, 3H).

[0192] Referring to the conditions and steps of the first synthetic method in Example 1, 3,4,5-trifluoroaniline was reacted with compound 5 to obtain 3-(benzyloxy)-N-(3,4,5-trifluorophenyl)thiophene-2-carboxamide (Formula 12-10) with a yield of 50%. MS (ESI) (m / z): 364 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.24 (s, 1H), 7.55 (m, 5H), 7.32 (s, 1H), 7.06 (d, J = 21.9Hz, 3H), 5.44 (s, 2H).

[0193] Referring to the conditions and steps of the first synthetic method in Example 1, 4-aminopyridine was reacted with compound 5 to obtain 3-(benzyloxy)-N-(pyridin-4-yl)thiophene-2-carboxamide (Formula 12-11) with a yield of 97%. MS (ESI) (m / z): 311 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.30(s,1H),8.41(m,2H),7.50(m,5H),7.26(m,3H),6.99(s,1H),5.28(s,2H).

[0194] Referring to the conditions and steps of the first synthetic method in Example 1, 2-amino-6-methylpyridine was reacted with compound 5 to obtain 3-(benzyloxy)-N-(6-methylpyridin-3-yl)thiophene-2-carboxamide (Formula 12-12) with a yield of 94%. MS (ESI) (m / z): 325 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.38(s,1H),8.43(s,1H),7.91(d,J=8.1Hz,1H),7.85(s,1H),7 .58(m,2H),7.44(s,3H),7.28(s,1H),7.22(d,J=8.7Hz,1H),5.42(s,2H),2.41(s,3H).

[0195] Referring to the conditions and steps of the first synthetic method in Example 1, 3-amino-6-fluoropyridine was reacted with compound 5 to obtain 3-(benzyloxy)-N-(6-fluoropyridin-3-yl)thiophene-2-carboxamide (Formula 12-13) with a yield of 20%. MS (ESI) (m / z): 329 [M+H] + . 1 H NMR (400MHz, CDCl3) δ9.23 (s, 1H), 8.26 (s, 1H), 7.85 (s, 1H), 7.51 (d, J = 19.4Hz, 5H), 7.27 (s, 1H), 7.00 (s, 1H), 6.88 (s, 1H), 5.28 (s, 2H).

[0196] Referring to the conditions and steps of the first synthetic method in Example 1, 3-amino-6-trifluoromethylpyridine was reacted with compound 5 to obtain 3-(benzyloxy)-N-(6-trifluoromethylpyridin-3-yl)thiophene-2-carboxamide (Formula 12-14) with a yield of 48%. MS (ESI) (m / z): 379 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.76 (s, 1H), 8.73 (s, 1H), 8.34 (d, J = 7.4Hz, 1H), 7.90 (m, 2H),7.59(d,J=6.4Hz,2H),7.47–7.37(m,3H),7.28(d,J=5.2Hz,1H),5.45(s,2H).

[0197] Referring to the conditions and steps of the first synthetic method in Example 1, 3-benzyloxypyridine-2-carboxylic acid was reacted with compound 6 to obtain 3-benzyloxy-N-(pyridin-3-yl)pyridine-2-carboxamide (Formula 12-15) with a yield of 43%. MS (ESI) (m / z): 306 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.67(s,1H),8.87(d,J=2.5Hz,1H),8.28(dd,J=4.7,1.5Hz,1H),8.26–8.21(m,1H),8.18(d,J=8.1Hz,1 H),7.75–7.67(m,1H),7.54(dd,J=8.5,4.6Hz,1H),7.46(d,J=7.2Hz,2H),7.41–7.31(m,3H),7.29(d,J=7.2Hz,1H),5.25(s,2H).

[0198] Referring to the conditions and steps of the first synthetic method in Example 1, 4-benzyloxypyridine-3-carboxylic acid was reacted with compound 6 to obtain 4-benzyloxy-N-(pyridin-3-yl)pyridine-3-carboxamide (Formula 12-16) with a yield of 30%. MS (ESI) (m / z): 306 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.33(s,1H),8.61(d,J=2.4Hz,1H),8.53(s,1H),8.45(d,J=5.8Hz,1H),8.16(d,J=4.8Hz,1H), 7.99(d,J=8.4Hz,1H),7.39(d,J=7.3Hz,1H),7.31(t,J=6.0Hz,1H),7.26–7.18(m,4H),7.07–7.01(m,1H),5.19(s,2H).

[0199] Referring to the conditions and steps of the first synthetic method in Example 1, 3-benzyloxypyridine-4-carboxylic acid was reacted with compound 6 to obtain 3-benzyloxy-N-(pyridin-3-yl)pyridine-4-carboxamide (Formula 12-17) with a yield of 40%. MS (ESI) (m / z): 306 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.58(s,1H),8.69(d,J=34.6Hz,2H),8.32(dd,J=22.3,4.6Hz,2H),8 .09(d,J=8.2Hz,1H),7.52(dd,J=28.8,5.9Hz,3H),7.35(dd,J=16.5,9.5Hz,4H),5.34(s,2H).

[0200] Referring to the conditions and steps of the first synthetic method in Example 1, 2-benzyloxypyridine-3-carboxylic acid was reacted with compound 6 to obtain 2-benzyloxy-N-(pyridin-3-yl)pyridine-3-carboxamide (Formula 12-18) with a yield of 36%. MS (ESI) (m / z): 306 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ10.46(s,1H),8.74(d,J=2.6Hz,1H),8.42–8.34(m,1H),8.31(d,J=4.7Hz,1H),8 .12(d,J=7.3Hz,2H),7.54(d,J=7.2Hz,2H),7.43–7.28(m,4H),7.21(dd,J=7.3,4.9Hz,1H),5.49(s,2H).

[0201] Referring to the conditions and steps of the first synthetic method in Example 1, 4-benzyloxypyrimidine-5-carboxylic acid was reacted with compound 6 to obtain 4-benzyloxy-N-(pyridin-3-yl)pyrimidine-5-carboxamide (Formula 12-19) with a yield of 60%. MS (ESI) (m / z): 307 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.56(s,1H),8.95(s,1H),8.85(s,1H),8.75(d,J=2.5Hz,1H),8.30( dd,J=4.7,1.5Hz,1H),8.13–8.06(m,1H),7.55–7.45(m,2H),7.41–7.30(m,4H),5.53(s,2H).

[0202] Referring to the conditions and steps of the first synthetic method in Example 1, 3-benzyloxypiperazine-2-carboxylic acid was reacted with compound 6 to obtain 3-benzyloxy-N-(pyridin-3-yl)piperazine-2-carboxamide (Formula 12-20) with a yield of 60%. MS (ESI) (m / z): 307 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.73(s,1H),8.73(d,J=2.5Hz,1H),8.31(d,J=2.6Hz,1H),8.22(d,J=2.6Hz,1H),8.18(dd,J=4.7,1.4H z,1H),8.04(dd,J=8.5,1.9Hz,1H),7.34(d,J=7.2Hz,2H),7.30–7.25(m,1H),7.25–7.20(m,2H),7.19–7.14(m,1H),5.35(s,2H).

[0203] The structure of the comparative compound is as follows:

[0204]

[0205] Example 12: Determination of the inhibitory effect of alkoxythiophene carboxylic acid aromatic amide compounds (including carboxylic acid / phosphate compounds derived therefrom as prodrugs) on SMS2 in vitro

[0206] Experimental instruments and materials

[0207] 1. Electric constant temperature water bath (Shanghai Yiheng Technology Co., Ltd.).

[0208] 2. Vortex mixer (Shanghai Jingke Industrial Co., Ltd. Model XW-80A).

[0209] 3. High-speed centrifuge (model Eppendorf 5804R).

[0210] 4. High performance liquid chromatography Agilent 1100 (Agilent Technologies, Palo Alto, CA, USA) equipped with quaternary pump, vacuum degassing, and FLD fluorescence detector.

[0211] 5.HPLC column COSMOSIL 5C18-MS-II (4.6mm IDX 250mm).

[0212] 6. DMPC was purchased from Santa Cruz (USA) and dissolved in ethanol at a concentration of 40 mmol / L. C6-NBD-Ceramide was purchased from Santa Cruz (USA) and dissolved in DMSO at a concentration of 1.16 mmol / L.

[0213] 7. All organic solvents used were purchased from Shanghai Sinopharm Reagent Company. Methanol was chromatographic grade. Water was ultrapure water filtered by a Milli-Q pump, deionized, and ultrafiltered through a 0.22 μm membrane. Other biological consumables were purchased from domestic companies.

[0214] 8. Prepare the test compound solution: Accurately weigh 1-2 mg of each test compound, first add an appropriate amount of DMSO to accurately prepare a 10 mmol / L stock solution. Take a certain volume of the DMSO stock solution of the test compound, and then add an appropriate volume of DMSO to dilute the test compound to the desired concentration.

[0215] 9. SMS2 pure enzyme, DDM solution and buffer were provided by Cao Yu's research group at the National Center for Protein Science (Shanghai).

[0216] Detection of inhibitory activity of alkoxythiophene carboxylic acid aromatic amide compounds (including carboxylic acid / phosphate compounds derived therefrom as prodrugs) against SMS2

[0217] Take 6μL SMS2 protein (0.144mg / mL) and add it to 3mL DDM buffer solution for dilution and vortex. Take 79μL of the above enzyme dilution solution and add it to a 1.5mL EP tube, add 1μL of the inhibitor DMSO solution, and vortex for 15s. Let it stand for 5min at room temperature. Add 20μL of the mixed solution of DMPC and C6-NBD-ceramide and vortex for 15s. Incubate at 37℃ for 30min. Add 200μL of anhydrous ethanol to quench the reaction and vortex for 15s. Take out 200μL and store it in the dark at 4℃ for HPLC detection.

[0218] Reference (Xiaodong Deng, et al. Analytical Letters, 2012, 45(12): 1581-1589.), the same high performance liquid chromatography method as in the literature was used to perform fluorescence quantitative analysis of the samples prepared above. The peak area A(sm) value and A(ceramide) value of C6-NBD-SM and C6-NBD-Ceramide in the DMSO group and inhibitor group samples on the corresponding HPLC spectra were analyzed and recorded, and the inhibition rate of the test compound was calculated according to the following formula.

[0219]

[0220] The inhibition rate of the target compound (including the comparative example compound) on SMS2 at a concentration of 1 μM or 10 μM was measured as follows:

[0221]

[0222] (Each set of data was measured twice and the average value was taken; “-” means not measured).

[0223] Example 13: Evaluation of the therapeutic effect of SMS2 inhibitors on dry eye in mice

[0224] (1) Experimental Materials

[0225] Experimental animals: 6-8 week old male C57BL / 6 experimental mice.

[0226] Experimental instruments: Alisn constant temperature air drying oven, desktop microscope, cobalt blue lamp.

[0227] Experimental reagents and consumables: phenol red cotton thread, 1% sodium fluorescein solution, Xingqi eye drops 0.05% cyclosporine eye drops (Ⅱ) 0.04 mg / g of Example Compound 1-2 preparation, 0.04 mg / g of Example Compound 1-31 preparation.

[0228] (2) Experimental methods

[0229] Twenty-eight male C57BL / 6 experimental mice aged 6-8 weeks were placed in a constant temperature blast drying oven and controlled with an ambient humidity of 19.5% and an ambient temperature of 25°C for 2 weeks to establish a dry eye model. At the end of the 2nd week, the dry eye mice were randomly divided into 4 groups, with 7 mice in each group. Subsequently, the humidity of the constant temperature blast drying oven was adjusted to 50% and the mice were continued to be raised for one week. At the same time, each group of mice received corresponding drug treatment: the vehicle group was treated with an empty vehicle without SMS2 inhibitor; the cyclosporine group was treated with commercial cyclosporine eye drops; the Example compound 1-2 group was treated with 0.04% compound 1-2; and the Example compound 1-31 group was treated with 0.04% compound 1-31. Each eye of the mouse received 5μl of pharmaceutical ocular surface administration twice a day. Another 7 male C57BL / 6 experimental mice aged 6-8 weeks were raised in a constant temperature blast drying oven with an ambient humidity of 50% and an ambient temperature of 25°C throughout the whole process, and received an empty vehicle as a physiological control in the third week.

[0230] At the end of the third week, all mice were scored for ocular surface fluorescein sodium staining to assess the recovery of ocular surface damage, and the tear secretion of mice was measured with phenol red thread. The specific operation was as follows: the phenol red thread was placed in the conjunctival sac of the mouse under anesthesia for 30 seconds, and then the discoloration length of the phenol red thread was measured. Subsequently, 2 μl of 1% sodium fluorescein solution was given to the ocular surface, and photos were taken under cobalt blue light and scored and counted.

[0231] Experimental Results

[0232] The experimental results show that SMS2 inhibitors have significant therapeutic effects on the treatment of corneal epithelial damage caused by dry eyes, and can also promote tear secretion in dry eye mice. Two SMS2 inhibitors, Example Compound 1-2 and Example Compound 1-31, both showed therapeutic effects in the treatment of dry eyes ( Figure 1 and Figure 2 ), which is manifested by a decrease in corneal fluorescein sodium staining scores, and has a statistically significant advantage over the commercial cyclosporine eye drops commonly used in clinical dry eye. Figure 3 ), cyclosporine and two SMS2 inhibitors showed an upward trend compared with the vehicle group. These results suggest that SMS2 inhibitors have the potential to treat dry eye.

[0233] The above is only a preferred embodiment of the present invention. It should be pointed out that ordinary technicians in this technical field can make several improvements and modifications without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An alkoxythiophenecarboxylic acid aromatic amide compound of the structure represented by formula (1), or a pharmaceutically acceptable salt thereof: in, X, Y, and Z are each independently selected from a carbon atom or a sulfur atom; R 1 is selected from substituted or unsubstituted phenyl, pyridyl, pyridazinyl, pyrimidinyl, thiazolyl, The substituents of the substituted groups are selected from 1 to 5 of halogen, methyl, hydroxy, methoxy, trifluoromethyl, trifluoromethoxy, cyano, and nitro; R 2 Selected from hydrogen, alkyl with less than 3 carbon atoms, and cycloalkyl; R 3 is selected from substituted or unsubstituted hydrocarbon groups; R 4 One or two selected from hydrogen, alkyl with less than 3 carbon atoms, halogen, and nitro.

2. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The X and Y are each independently selected from a sulfur atom or a carbon atom, and the Z is a carbon atom.

3. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The R 3 Select one of the following structures: Wherein n is an integer from 1 to 10; W is selected from a carbon atom or a nitrogen atom; L and M are each independently selected from -C=C-, a sulfur atom, and an oxygen atom; R 5 1 to 5 selected from hydrogen, substituted or unsubstituted alkyl with less than 4 carbon atoms, substituted or unsubstituted alkoxy with less than 20 carbon atoms, halogen, acyl, nitro, cyano, hydroxyl, carboxylic (phosphonic) ester, and aromatic heterocycle.

4. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 3, or a pharmaceutically acceptable salt thereof, characterized in that: The alkoxy group is an alkoxy group with no more than 15 carbon atoms.

5. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The compound is:

6. The alkoxythiophene carboxylic acid aromatic amide compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The pharmaceutically acceptable salt is hydrochloride, hydrobromide, tartrate, methanesulfonate, sodium salt, potassium salt, calcium salt, ammonium salt or amino acid salt.

7. A pharmaceutical composition, characterized in that Contains at least one therapeutically effective amount of an alkoxythiophenecarboxylic acid aromatic amide compound represented by formula (1) as described in any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof.

8. Use of the alkoxythiophene carboxylic acid aromatic amide compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, in the preparation of a small molecule inhibitor of sphingomyelin synthase.

9. Use of the alkoxythiophene carboxylic acid aromatic amide compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for preventing and treating a disease caused by an abnormal increase in sphingomyelin levels.

10. The use of claim 9 in the preparation of a drug for preventing and treating a disease caused by an abnormal increase in sphingomyelin levels, characterized in that: The diseases caused by abnormal increase in sphingomyelin levels include atherosclerosis, obesity, insulin resistance, fatty liver, type II diabetes, enteritis, pulmonary edema, lung damage, malignant tumors, Alzheimer's disease and dry eye.

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